How to dilute loading dye? (For gel electrophoresis) - (Jun/18/2007 )

I have a quantity of loading dye (6X w/ 40% sucrose w/v) to be used in gel electrophoresis.

I have heard of people using anything from 1X to 3X concentration. Are there any special procedures to dilute loading dye?

-Cedricb-

use glycerol i guess

-Aris-

no. there is no dilution special procedures. It all depends on how intense and how dense you want your samples to be. I like my samples dense and strongly coloured. thus I use 4x. Another labmate uses 2x. The boss uses 1x. Personal perference

-perneseblue-

QUOTE (perneseblue @ Jun 18 2007, 07:01 PM)

no. there is no dilution special procedures. It all depends on how intense and how dense you want your samples to be. I like my samples dense and strongly coloured. thus I use 4x. Another labmate uses 2x. The boss uses 1x. Personal perference

Ok, got it. Do you just dilute it with dH2O?

-Cedricb-

QUOTE (Cedricb @ Jun 19 2007, 10:52 AM)

QUOTE (perneseblue @ Jun 18 2007, 07:01 PM)

no. there is no dilution special procedures. It all depends on how intense and how dense you want your samples to be. I like my samples dense and strongly coloured. thus I use 4x. Another labmate uses 2x. The boss uses 1x. Personal perference

Ok, got it. Do you just dilute it with dH2O?

You can dilute with dH2O but you can also just use with your sample proportionally. E.g. you have 20ul sample and want to run with 1x loading dye, then add 4ul of your 6xloading buffer into your sample and just run the total 24ul instead of diluding your LB to 2x and then use 20ul of dye with 20ul sample.

-Almasy-

[/quote]

You can dilute with dH2O but you can also just use with your sample proportionally. E.g. you have 20ul sample and want to run with 1x loading dye, then add 4ul of your 6xloading buffer into your sample and just run the total 24ul instead of diluding your LB to 2x and then use 20ul of dye with 20ul sample.[/quote]what LB stand for here???may be it would ne LD?? loading dye?? i think if i use 24 ul sample per well, my well will overflow and contaminate neighbour well

-T. reesei-

QUOTE (T. reesei @ Jun 22 2007, 06:16 AM)

what LB stand for here???may be it would ne LD?? loading dye?? i think if i use 24 ul sample per well, my well will overflow and contaminate neighbour well

LB is loading buffer in this case. If you use 24ul and it already overflows, think about the other choice, when you use 20ul of 2x dye and 20ul of sample (total 40ul).